Mice at 12 weeks old were housed in individual cages two days before the beginning of the experiment. Mice were deprived of food for 2 h after the dark period and i.p. injected with ghrelin (0.4 μg/g of body weight; Merck Millipore, Cat# 494127-100G) in physiological saline solution (B. Braun; Cat# 12260029_1019) at 0 min and again at 30 min. We monitored the eating time and food intake for 1 h after the first injection [44 (link)].
Physiological saline solution
Manufactured by B. Braun
Sourced in Germany
Physiological saline solution is a sterile, isotonic solution of sodium chloride in purified water. It is designed to closely match the composition of human body fluids, making it suitable for various medical and laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Ghrelin, by Merck Group (1 mentions)
Spss statistics 20, by IBM (1 mentions)
Protease inhibitor, by Merck Group (1 mentions)
Purelab flex 4, by Elga LabWater (1 mentions)
Maldi biotyper, by Bruker (1 mentions)
Methylene blue, by Carl Roth (1 mentions)
Sodium dodecyl sulfate (sds), by Carl Roth (1 mentions)
Halt protease inhibitor cocktail, by Merck Group (1 mentions)
5 protocols using physiological saline solution
1
Ghrelin Modulation of Feeding Behavior in Mice
2
Thermal Imaging of Animal Motor Activity
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We studied motor activity in 15 animals (8 males, 7 females) by recording 30-min videos with a thermal camera (ThermaCAM SC2000 PAL, Flir Systems), under light (780 lx) or dark (0 lx) conditions. The two videos were recorded with a 48-h interval between videos. These animals were subjected to the same pre-imaging animal handling by anaesthetizing mice with inhalation anaesthesia (3% sevoflurane in 100% oxygen), then administering an intravenous injection of 0.2 ml physiological saline solution (0.9% NaCl, B Braun Medical). The recorded videos were analysed by an expert, who timed the periods of animal activity (expressed in seconds). We defined activity as animal movement over the cage floor or its grid. The first 2-min of the video were discarded, because they corresponded to recovery from the inhaled anaesthesia. An ANOVA linear mixed model was used to analyse the differences in motor activity between light and dark environments (IBM SPSS Statistics 20). Sex, the illumination condition, and the interaction between the two were fixed effects, and the individual mouse was the repeated factor. Data are expressed as the mean ± standard deviation.
3
Saliva Collection for Stem Cell Patients
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Sample collection was based on the protocols described previously14 (link),15 . In brief, following oral cavity rinse with 25 ml of physiological saline solution (B. Braun Melsungen AG, Melsungen, Germany) for 30 s, saliva was collected for 5 min in 15 ml disposable, pre-disinfected tubes (Sigma-Aldrich, St. Louis, MO, USA). Participants adapted to the condition for 5 min. Taking into account the diurnal variation of saliva constituents, samplings were done every morning at the same time, 1 h after eating, drinking or toothbrushing. Patients in the Haematopoietic Stem Cell Transplantation Centre’s steril rooms used a steril disposable oral swab (DenTips MDS096502, Medline Industries.Inc., Mundelein, IL, USA) impregnated with physiological saline solution to maintain oral hygiene during the period of cytopenia. Protease Inhibitor (Sigma-Aldrich, St. Louis, MO, USA) was added to the collected saliva samples and after aliquoting into 1.5 ml Eppendorf tubes, samples were stored at − 70 °C until processing.
4
Synthesis of Bolalipid Compounds
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The bolalipids dotriacontane-1,1′-diylbis [2-(trimethylammino)ethyl phosphate] (PC-C32-PC) and dotriacontane-1,1′-diylbis [2-(dimethylammino)ethyl phosphate] (Me2PE-C32-Me2PE) were synthesized at the MLU Halle-Wittenberg (Halle, Germany) according to procedures described previously [40 (link),41 (link)]. Sodium dodecyl sulfate and methylene blue were purchased from Carl Roth GmbH and Co. KG (Karlsruhe, Germany). Hydroxyethyl cellulose 300 (HEC 300; Mw ~ 807 g/mol) was obtained from Caesar and Loretz GmbH (Hilden, Germany). Physiological saline solution was purchased from B Braun (Melsungen, Germany). Saccharomyces cerevisiae were obtained from a retail outlet. Clinically isolated Staphylococcus aureus (ATCC 25923) was determined using MALDI Biotyper (Bruker Corporation, Billerica, MA, USA). Fertilized chicken eggs were obtained from Brormann GmbH (Rheda-Wiedenbrück, Germany). For all experiments ultrapure water from PURELAB®® flex 4 (ELGA LabWater, High Wycombe, UK) was used.
5
Standardized Saliva Collection for Biomarker Analysis
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Saliva collection was performed according to the standard methods [12 (link)]. Both controls and patients were in a sitting position during the sampling with eyes open and a slightly tilted head. Following oral cavity rinse with 25 mL of physiological saline solution (B. Braun Melsungen AG, Melsungen, Germany) for 30 s, saliva was collected for 5 min in an externally pre-disinfected 15 mL lockable Falcon tube (Sigma-Aldrich, St. Louis, MO, USA). Participants adapted to the test condition for 5 min prior to sample collection. Taking into account the diurnal variation of saliva constituents, samplings were done at a specified time window: between 7 a.m. and 8 a.m., one hour after eating, drinking, or tooth-brushing in order to avoid contamination. Patients in sterile rooms used a gauze plate or DenTips (MDS096502, Medline Industries. Inc., Mundelein, IL, USA), and a disposable oral swab, impregnated with physiological saline solution, in order to maintain optimal oral hygiene during the period of cytopenia. Within one hour of collection, Halt Protease Inhibitor Cocktail (Sigma-Aldrich, St. Louis, MO, USA) was added proportionally to the saliva samples. After homogenization, saliva samples were aliquoted into 1.5 mL Eppendorf tubes and stored at −70 °C until further processing.
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